Molecular cloning and bioinformatics analyses of a GH3 beta-glucosidase from Auricularia heimuer

Figures & data

Table 1. Primers used in this study.

Primer	Sequence (5^,–3^,)
A-bgl	5^, -ATGGCGCCGTCCGACTTTGC-3^,
A-bglR	5^, -CTACAACCCGGTCCAGAGGACGTCC-3^,
A-bglhF	5^, -GCCATGGCTGATATCGGATCCATG GCGCCGTCCGACTTTGC-3^,
A-bglhR	5^, -TCGAGTGCGGCCGCAAGCTTCTACAA CCCGGTCCAGAGGACGTCC-3^,
pET-F	5^, -TAATACGACTCACTATAGGG-3^,
pET-R	5^, -GCTAGTTATTGCTCAGCGG-3^,
Aa-bglqF	5^, -GGCTTCGGTTTCTCATACAC-3^,
Aa-bglqR	5^, -CCAGGCTTCAAGTCCTCTAC-3^,
Actin-F	5^, -GCCCTCGTTCCTTGGTCTTG-3^,
Actin-R	5^, -GTGCTCAACGACGCCAGGAT-3^,

The underscore represents a homologous sequence.

Figure 1. Different stages of fruiting bodies of Auricularia heimuer. A, B and C represent primordium (A), immature fruiting bodies (B) and mature fruiting bodies (C) in cultivation bags, while a, b and c indicate materials collected from the corresponding cultivation bags.

Figure 2. Nucleotide sequence and deduced amino acid sequence of Aa-bgl.

Figure 3. Amino acid content of the Aa-bgl protein.

Figure 4. Conserved domains in the Aa-bgl protein.

Figure 5. Transmembrane region prediction for the Aa-bgl protein.

Figure 6. Signal peptide prediction for the Aa-bgl protein.

Figure 7. Structural and expression analysis of the Aa-bgl protein. (a) Tertiary structure prediction. (b) SDS-PAGE analysis following induction of expression by IPTG (1 mmol/L). M, protein molecular weight markers (Solarbio); Lane 1, E. coli transformed with pET-32a; Lanes 2–7, E. coli transformed with pET-32a-bgl induced with IPTG for 0, 2, 4, 6, 8 and 10 h, respectively.

Figure 8. Phylogenetic tree of A. heimuer Aa-bgl and homologs from various species.

Table 2. Sequences of Aa-bgl homologs in various species.

The source species	Accession number
Lentinula edodes	GAW03012.1
Lentinula edodes	GAW03013.1
Gymnopus luxurians	KIK64526.1
Heliocybe sulcata	TFK45937.1
Dentipellis sp.	KAA1466649.1
Rickenella mellea	TDL18908.1
Schizopora paradoxa	KLO19583.1
Fomitiporia mediterranea	XP 007264285.1
Sanghuangporus baumii	OCB89642.1
Inonotus obliquus	AMK37826.1
Sistotremastrum suecicum	KZT33940.1
Auricularia subglabra	EJD40098.1
Exidia glandulosa	KZV82289.1
Exidia glandulosa	KZV96315.1
Auricularia subglabra	EJD35757.1
Auricularia subglabra	EJD37897.1
Pseudothermotoga thermarum	AEH51751.1

Figure 9. Aa-bgl transcription levels at different stages of A. heimuer development.

Note: Values are means ± standard deviation from three experiments.

Data availability statement

The data that support the findings of this study are available on request from the corresponding author, [Z]. The data are not publicly available due to their containing information that could compromise the privacy of research participants.